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Fitc and tritc filters

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FITC and TRITC filters are optical filters used in fluorescence microscopy. FITC (Fluorescein Isothiocyanate) filters are designed to isolate the green emission of fluorescein-labeled samples, while TRITC (Tetramethylrhodamine Isothiocyanate) filters are used to isolate the red emission of rhodamine-labeled samples. These filters allow for the selective visualization of specific fluorophores in a sample.

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Lab products found in correlation

Vectashield containing dapi, by Vector Laboratories (4 mentions) Eclipse e400 microscope, by Nikon (4 mentions) Alexafluor 488 conjugated goat anti mouse igg1, by Thermo Fisher Scientific (3 mentions) Rabbit anti hcd144, by Cell Signaling Technology (2 mentions) Rabbit anti hcd133, by Miltenyi Biotec (2 mentions) Rabbit anti hcd271, by Alomone (2 mentions) Rat anti mcd31, by BD (2 mentions) Alexa fluor 594 conjugated donkey anti rat igg, by Thermo Fisher Scientific (2 mentions) Fitc conjugated donkey anti rabbit igg, by Thermo Fisher Scientific (1 mentions) Alexa fluor 594 conjugated donkey anti rabbit igg, by Thermo Fisher Scientific (1 mentions) Rat anti mouse cd31, by BD (1 mentions)

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FITC filter

5 protocols using fitc and tritc filters

1

Immunofluorescence Staining of Corin-Treated Cells

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Glass coverslips were placed into 6-well plates, and 50,000 cells were plated in each well. Cells were treated with DMSO control or 100 nM or 1 μM corin for 72 hours. Media were aspirated and cells were fixed to slides with 4% paraformaldehyde in PBS for 10 minutes. Slides were then washed twice with 0.1% Triton X-100 in PBS. Blocking solution (10% goat serum in PBS with 0.25% Triton X-100) was added for 30 minutes, followed by primary antibodies (Supplemental Table 13) diluted in blocking solution at 1:500, incubated at 4°C overnight. The next day, slides were washed twice with 0.1% Triton X-100 in PBS. Secondary antibodies were FITC-conjugated donkey anti–rabbit IgG, Alexa Fluor 594–conjugated donkey anti–rabbit IgG, and Alexa Fluor 488–conjugated goat anti–mouse IgG1 (Invitrogen, Thermo Fisher Scientific). Isotype-matched rabbit, mouse, or rat Ig was used in place of the primary antibody for the control. Sections were mounted with VECTASHIELD containing DAPI (H-1800, Vector Laboratories) and examined under a Nikon Eclipse E400 microscope equipped with FITC and TRITC filters (Nikon) and a Mercury 100W lamp (Chiu Technical Corp.).
Wu M., Hanly A., Gibson F., Fisher R., Rogers S., Park K., Zuger A., Kuang K., Kalin J.H., Nocco S., Cole M., Xiao A., Agus F., Labadorf A., Beck S., Collard M., Cole P.A, & Alani R.M. (2024). The CoREST repressor complex mediates phenotype switching and therapy resistance in melanoma. The Journal of Clinical Investigation, 134(6), e171063.
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2

Quantifying Melanoma Angiogenesis and Vasculogenic Mimicry

Cited 1 time
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Frozen melanoma xenograft sections were subjected to double immunofluorescence using standard procedures (29 (link)). The primary antibodies used in this study are rabbit anti-hCD144 (human-specific; Cell Signaling Technology), rabbit anti-hCD133 (human-specific; Miltenyi Biotech Inc.), rabbit anti-hCD271 (human-specific; Alomone Labs) and rat anti-mCD31 (mouse-specific; BD Biosciences). The secondary antibodies employed are FITC-conjugated donkey anti-rabbit IgG, AlexaFluor 594-conjugated donkey anti-rat IgG, and AlexaFluor 488-conjugated goat anti-mouse IgG1 (Invitrogen) antibodies. Isotype-matched rabbit, mouse or rat immunoglobulin was used in place of the primary antibody for control. Sections were mounted with VectaShield containing DAPI (Vector Laboratories, Burlingame, CA) and examined under a Nikon Eclipse E400 microscope equipped with FITC and TRITC filters (Nikon) and a Mercury-100W lamp (Chiu Technical Corporation). Host angiogenesis (mCD31+) was quantified by measuring luminal area or number of tubules per field (40 ×) using ImageJ software, and data were analyzed using the Student t test. Quantification of human melanoma–derived VM channels was performed by measuring the hCD144+ area per field (100 ×) using ImageJ software, and data were analyzed using the Student t test.
Hsu M.Y., Yang M.H., Schnegg C.I., Hwang S., Ryu B, & Alani R.M. (2017). Notch3 Signaling-Mediated Melanoma-Endothelial Crosstalk Regulates Melanoma Stem-Like Cell Homeostasis and Niche Morphogenesis. Laboratory investigation; a journal of technical methods and pathology, 97(6), 725-736.
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3

Melanoma Xenograft Angiogenesis and Vasculogenic Mimicry

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Consecutive frozen melanoma xenograft sections were subjected to double indirect immunofluorescence according to standard protocols. The primary antibodies used were rabbit anti-human CD144 (Cell Signaling Technology) at 1:100, biotinylated anti-human CD133 (Miltenyl Biotec Inc.) at 1:20, rabbit anti-human CD271 (Alomone) at 1:100, and rat anti-mouse CD31 (BD Biosciences, San Jose, CA) at 1:50. The secondary antibodies used were FITC-conjugated donkey anti-rabbit IgG (Accurate Chemical & Scientific Corporation, Westbury, NY) at 1:100, FITC-conjugated mouse anti-biotin IgG (Jackson Immunoresearch) at 1:100, and TRITC-goat anti-rat igG (Jackson Immunoresearch) at 1:100. For control, isotype-matched immunoglobins (Vector Laboratories, Burlingame, CA) were used in place of the primary antibody. Sections were mounted with Vectashield containing DAPI (Vector Laboratories) and examined under a Nikon Eclipse E400 microscope (Melville, NY) equipped with FITC and TRITC filters (Nikon, Tokyo, Japan) and a Mercury-100W lamp (Chiu Technical Corporation, Kings Park, NY). Host angiogenesis (mCD31+) and human melanoma-derived VM channels (hCD144+) were quantified as measured by luminal area per field (100×) using Image J software, and data was analyzed using the Student t test.
Schnegg C.I., Yang M.H., Ghosh S.K, & Hsu M.Y. (2015). Induction of vasculogenic mimicry overrides VEGF-A silencing and enriches stem-like cancer cells in melanoma. Cancer research, 75(8), 1682-1690.
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4

Competition Assay for VHH Epitope Mapping

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Competition experiments were designed to determine whether two unique VHHs compete for the same epitope. For this application, VHHs were expressed with different epitope tags, specifically myc and E-tag. Briefly, sporozoite samples were prepared on glass slides, fixed, dried and blocked as described above. Sporozoites were then probed simultaneously with two unique VHHs, each with a different tag, such that the concentration of a myc-tagged VHH exceeded that of an E-tagged VHH by 10-fold, and vice versa. Binding of an E-tagged VHH was detected with anti-E-tag FITC conjugate at 1:100 dilution. Binding of a myc-tagged VHH was detected by subsequent probing with the 9E10 anti-myc mAb at 10 μg/mL, followed by anti-mouse IgG Alexa Fluor 568 Ab at 1:500 dilution. Same frame fluorescent micrographs were obtained at constant exposure using FITC and TRITC filters (Nikon Instruments Inc). Corresponding images were color-merged using ImageJ 1.48v software (Rasband, 2018).
Jaskiewicz J.J., Tremblay J.M., Tzipori S, & Shoemaker C.B. (2021). Identification and characterization of a new 34 kDa MORN motif-containing sporozoite surface-exposed protein, Cp-P34, unique to Cryptosporidium. International journal for parasitology, 51(9), 761-775.
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5

Quantifying Melanoma Angiogenesis and Vasculogenic Mimicry

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Frozen melanoma xenograft sections were subjected to double immunofluorescence using standard procedures (29 (link)). The primary antibodies used in this study are rabbit anti-hCD144 (human-specific; Cell Signaling Technology), rabbit anti-hCD133 (human-specific; Miltenyi Biotech Inc.), rabbit anti-hCD271 (human-specific; Alomone Labs) and rat anti-mCD31 (mouse-specific; BD Biosciences). The secondary antibodies employed are FITC-conjugated donkey anti-rabbit IgG, AlexaFluor 594-conjugated donkey anti-rat IgG, and AlexaFluor 488-conjugated goat anti-mouse IgG1 (Invitrogen) antibodies. Isotype-matched rabbit, mouse or rat immunoglobulin was used in place of the primary antibody for control. Sections were mounted with VectaShield containing DAPI (Vector Laboratories, Burlingame, CA) and examined under a Nikon Eclipse E400 microscope equipped with FITC and TRITC filters (Nikon) and a Mercury-100W lamp (Chiu Technical Corporation). Host angiogenesis (mCD31+) was quantified by measuring luminal area or number of tubules per field (40 ×) using ImageJ software, and data were analyzed using the Student t test. Quantification of human melanoma–derived VM channels was performed by measuring the hCD144+ area per field (100 ×) using ImageJ software, and data were analyzed using the Student t test.
Hsu M.Y., Yang M.H., Schnegg C.I., Hwang S., Ryu B, & Alani R.M. (2017). Notch3 Signaling-Mediated Melanoma-Endothelial Crosstalk Regulates Melanoma Stem-Like Cell Homeostasis and Niche Morphogenesis. Laboratory investigation; a journal of technical methods and pathology, 97(6), 725-736.
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